Study On The Technique System About Tissue Culture And Rapid Propagation Of Cymbidium Grandiflorium

This research takes root stumps as explant and adopts the regeneration way of PLB to establish a rapid technique system of micropropagation in Cymbidium Grandiflorium.This research resolves around:exterminating germs of explants,PLB inducement,multiplication of PLB and shoot clumps,sound seedling and rooting and transplanting finally.The experiment mainly focus on the following aspects:the selection of the best explant asepsis method,basic mediums and raising mode,the combination and concentration of hormone and the selection of the best natural addiction,carbon source.Orthogonal Design was adopted in the experiments,all data analysis was dealt with by procedure SAS.The effected factors and the most suitable medium in each micropropagation stage were researched fully,then the technique parameter combination was filtrated,the superior regeneration system about Cymbidium Grandiflorium was to built up to improve the propagation coefficient,and,thus enhancing Cymbidium Grandiflorium commercial production efficiency,provide solution to the current market shortage of fine breed Cymbidium Grandiflorium,and technical support for professional nursery.The results were as follows:The effective sterilization method which takes root stumps as explant is:root stumps→washed by running water→dipped in the detergent water about 10min and used a soft brush to clean surface of buds→washed about 30min by running water→putted on the extra-clean desk→dipped in the 70%alcohol about 20s→washed 2 times by asepsis water→dipped in the 0.1%HgCl₂ 10min→washed 2 times by asepsis water→dipped in the 0.1%cephalosporin 5min→washed by asepsis water 2 times→2.0%NaClO2min→washed by asepsis water 5 times→water dried→inoculated.The significant factors of inducing PLB is BA and natural additions,and NAA is the subordination factor.The inducement medium of PLBs is:MS+BA4.0mg/L+ NAA0.2mg/L+ coconut milk 15%+ sugar30g/L+ agar8g/L+active cabon 1.0mg/L, pH5.4.The best multiplication medium of PLBs is:MS+BA2.0mg/L+ NAA0.2mg/L+ KT2.0mg/L+banana juice 100g/L+active cabon 1.0g/L+agar8g/L+sugar30g/L+PYP 200mg/L.The main factors of PLBs' differentiation are the basic medium,BA and NAA. The subordination factor is KT.And the best differentiation medium of PLB is:2MS +BA2.0mg/L+NAA0.1mg/L+KT3.0mg/L+banana juice100g/L+active cabon 1.0 g/L+agar8g/L+sugar30g/L+PVP200mg/L.The main factors of bud lumps multiplication are BA,the natural addictions,the height of seedling,the mode of seedling and the effect of carbon source.The seedlings about 0.5cm cut vertically at the bottom can obviously improve the multiplication of bud clumps.Banana juice 100mg/L and 4 seedlings in every 150ml bottle also obviously improve the multiplication of bud clumps.Sugar is the effective carbon source.The best multiplication medium is B5+KT 10mg/L+2, 4-D 0.2mg/L+banana juicel50g/L+citric acid 100mg/L+sugar 30g/L+agar 8.0g/L,pH5.1-5.4.The best multiplication medium of shoot clumps is: MS+BA2.0mg/L+NAA0.2mg/L+KT2.0mg/L+banana juice 100g/L+ active cabon 1.0 g/L+ agar 8g/L+ sugar 30g/L+PVP200mg/L.The best medium for promoting seedlings and rooting is 1/2MS + BA2.0 mg/L+NAA0.2mg/L+banana juicel00g/L+active cabon 1.0g/L+sugar 30g/L+ agar 8g/L,pH 5.4.